Širenje gena za rezistenciju na cefalosporine 3. generacije između bakterija bolničkih otpadnih voda

master's thesis

• Title (english): Spreading of the 3rd generation cephalosporins resistance genes among bacteria of hospital wastewater
• Author: Petra Gulan
• Mentor: Jasna Novak (mentor)
• Committee member: Tonči Rezić (predsjednik povjerenstva)
• Committee member: Nikolina Udiković Kolić (član povjerenstva)
• Committee member: Ana Bielen (član povjerenstva)
• Granter: University of Zagreb; Faculty of Food Technology and Biotechnology; (Department of Biochemical Engineering); (Laboratory for Antibiotic, Enzyme, Probiotic and Starter Cultures Technology); Zagreb
• Defense date and country: 2022-07-15, Croatia
• Scientific / art field, discipline and subdiscipline: BIOTECHNICAL SCIENCES; Biotechnology

Abstract

Antibiotička rezistencija predstavlja ozbiljan globalni problem zbog ograničenog izbora antibiotika za liječenje bakterijskih infekcija. Plazmidi su presudne genetičke determinante za širenje antibiotičke rezistencije. Upravo u ovom radu analiziran je plazmidni prijenos gena za rezistenciju na cefalosporine 3. generacije iz bakterija bolničkih otpadnih voda u Escherichia coli CV601. Egzogenom izolacijom plazmida dobiveno je 11 transkonjuganata s višestruko rezistentnim fenotipom, potvrđenim metodama disk difuzije i mikrodilucije. PCR analizom ustanovljeno je da plazmidi transkonjuganata sadrže minimalno jedan gen za β-laktamazu proširenog spektra (ESBL) pri čemu prevladava blaCTX-M‐ ₁. Uz ESBL gen(e), na 3 plazmida detektirana je prisutnost gena za karbapenemazu blaVIM. Na temelju PBRT analize plazmidi su svrstani u 4 skupine inkompatibilnosti pri čemu je dominirala IncL/M skupina. Budući da plazmidi ove skupine imaju širok raspon bakterijskih domaćina i nose višestruku rezistenciju, postoji rizik da upravo bolničke otpadne vode predstavljaju izvor i put daljnjeg širenja višestruko rezistentnih ESBL-producirajućih bakterija, uključujući patogene.

Abstract (english)

Antibiotic resistance presents a serious public health issue, due to reduced effective therapeutic approaches for the treatment of bacterial infections. Plasmids are key genetic determinants for the spread of antibiotic resistance. In this work, the plasmid transfer of genes from hospital wastewater bacteria resistant towards 3rd generation cephalosporins into Escherichia coli CV601 was analyzed. 11 transconjugants with a multiple resistance phenotype were obtained by exogenous isolation of the plasmid, which were confirmed by BOX-PCR. PCR analysis revealed that the plasmids of the transconjugants contain at least one extended-spectrum β-lactamase (ESBL) gene, with the highest frequency of blaCTX-M‐ ₁. Additionally, the carbapenemase genes (blaVIM), were detected on 3 plasmids. Based on PBRT analysis, plasmids were grouped into 4 groups of incompatibility with the IncL/M group prevailing. Since the plasmids of this group have a range of bacterial hosts and carry multiple resistances there is a risk that hospital wastewater can be a source of multiple resistant bacteria, including pathogens, and thus the risk of further spreading into the environment.

• Keywords: antibiotička rezistencija; BOX-PCR; egzogena izolacija plazmida; ESBL i karbapenemazni geni; PBRT analiza
• Keywords (english): antibiotic resistance; BOX-PCR; exogenous plasmid isolation; ESBL and carbapenemase genes; PBRT analysis
• Language: croatian
• URN:NBN: urn:nbn:hr:159:852816
• Study programme: Title: Molecular Biotechnology; Study programme type: university; Study level: graduate; Academic / professional title: magistar/magistra inženjer/inženjerka molekularne biotehnologije (magistar/magistra inženjer/inženjerka molekularne biotehnologije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2024-04-23 11:19:21